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. 2015 Jan 21;6(8):6014–6028. doi: 10.18632/oncotarget.3338

Figure 1. miR-1236 directly down-regulates the expression of AFP.

Figure 1

(A) The expression levels of miR-1236 (Left) and AFP (Right) mRNA in 20 pairs of HCC tissues and adjacent non-tumor tissues were detected by qRT-PCR. U6 and β-actin were used as internal controls to normalize the levels of miR-1236 and AFP, respectively. (B) qRT-PCR was used to detect the expression levels of miR-1236 (Left) and AFP (Right) mRNA in LO2, HepG2, Huh-7, SMMC-7721 and QGY-7703 cells. U6 and β-actin were used as internal controls to normalize the levels of miR-1236 and AFP, respectively. (C) The predicted binding sites for miR-1236 in the 3′UTR of AFP and the mutations in the binding sites are shown. (D) The EGFP reporter assay was performed in QGY-7703 cells cotransfected pcDNA3/EGFP-AFP 3′UTR wild type or pcDNA3/EGFP-AFP 3′UTRmut with pri-miR-1236 or ASO-miR-1236. (E) qRT-PCR was performed to detect the AFP mRNA level in QGY-7703 cells transfected with pcDNA3/pri-miR-1236, ASO-miR-1236 or the corresponding controls. (F) Western blot assays were used to detect the AFP protein level in QGY-7703 cells transfected with pcDNA3/pri-miR-1236, ASO-miR-1236 or the corresponding controls (Left), and the quantification of the bars are shown on the right. *p<0.05, **p<0.01, *** p<0.001, ns, no significance. All error bars indicate the means±SDs. All experiments were repeated at least three times.